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The cell adhesion molecule "CAR" and sialic acid on human erythrocytes influence adenovirus in vivo biodistribution.

Seiradake E, Henaff D, Wodrich H, Billet O, Perreau M, Hippert C, Mennechet F, Schoehn G, Lortat-Jacob H, Dreja H, Ibanes S, Kalatzis V, Wang JP, Finberg RW, Cusack S, Kremer EJ - PLoS Pathog. (2009)

Bottom Line: Using molecular, biochemical, structural and transgenic animal-based analyses, we found that the primary erythrocyte interaction domain for HAd37 is its sialic acid binding site, while CAV-2 binding depends on at least three factors: electrostatic interactions, sialic acid binding and, unexpectedly, binding to the coxsackievirus and adenovirus receptor (CAR) on human erythrocytes.We show that the presence of CAR on erythrocytes leads to prolonged in vivo blood half-life and significantly reduced liver infection when a CAR-tropic Ad is injected intravenously.This study provides i) a molecular and structural rationale for Ad-erythrocyte interactions, ii) a basis to improve vector-mediated gene transfer and iii) a mechanism that may explain the biodistribution and pathogenic inconsistencies found between human and animal models.

View Article: PubMed Central - PubMed

Affiliation: European Molecular Biology Laboratory, Grenoble Outstation, Grenoble, France.

ABSTRACT
Although it has been known for 50 years that adenoviruses (Ads) interact with erythrocytes ex vivo, the molecular and structural basis for this interaction, which has been serendipitously exploited for diagnostic tests, is unknown. In this study, we characterized the interaction between erythrocytes and unrelated Ad serotypes, human 5 (HAd5) and 37 (HAd37), and canine 2 (CAV-2). While these serotypes agglutinate human erythrocytes, they use different receptors, have different tropisms and/or infect different species. Using molecular, biochemical, structural and transgenic animal-based analyses, we found that the primary erythrocyte interaction domain for HAd37 is its sialic acid binding site, while CAV-2 binding depends on at least three factors: electrostatic interactions, sialic acid binding and, unexpectedly, binding to the coxsackievirus and adenovirus receptor (CAR) on human erythrocytes. We show that the presence of CAR on erythrocytes leads to prolonged in vivo blood half-life and significantly reduced liver infection when a CAR-tropic Ad is injected intravenously. This study provides i) a molecular and structural rationale for Ad-erythrocyte interactions, ii) a basis to improve vector-mediated gene transfer and iii) a mechanism that may explain the biodistribution and pathogenic inconsistencies found between human and animal models.

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Structure of the CAV-2 fiber head in complex with sialic acid and CAR.A. Ribbon diagram of CAV-2 fiber head in complex with sialyl-D-lactose (red = Neu5AC, yellow = fiber head). B. Cartoon representation of HAd37 fiber head in complex with sialyl-D-lactose (red = sialic acid, blue = fiber head). C. Surface charge representation of CAV-2 fiber head and cartoon representation of sialic acid as found in the complex. D. Surface charge representation of HAd37 fiber head and cartoon representation of sialic acid as found in the complex. E. Representation of the contacts between CAV-2 fiber head (purple) and sialic acid (yellow). F. Representation of the contacts between HAd37 fiber head (purple) and Neu5AC (yellow). G. Ribbon diagram of CAV-2 fiber head in complex with CAR D1 and sialyl-D-lactose (red = Neu5AC, green = CAR, yellow = fiber head). H. Ribbon diagram of HAd37 fiber head in complex with CAR and sialyl-D-lactose (red = Neu5AC, green = CAR, blue = fiber head).
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ppat-1000277-g003: Structure of the CAV-2 fiber head in complex with sialic acid and CAR.A. Ribbon diagram of CAV-2 fiber head in complex with sialyl-D-lactose (red = Neu5AC, yellow = fiber head). B. Cartoon representation of HAd37 fiber head in complex with sialyl-D-lactose (red = sialic acid, blue = fiber head). C. Surface charge representation of CAV-2 fiber head and cartoon representation of sialic acid as found in the complex. D. Surface charge representation of HAd37 fiber head and cartoon representation of sialic acid as found in the complex. E. Representation of the contacts between CAV-2 fiber head (purple) and sialic acid (yellow). F. Representation of the contacts between HAd37 fiber head (purple) and Neu5AC (yellow). G. Ribbon diagram of CAV-2 fiber head in complex with CAR D1 and sialyl-D-lactose (red = Neu5AC, green = CAR, yellow = fiber head). H. Ribbon diagram of HAd37 fiber head in complex with CAR and sialyl-D-lactose (red = Neu5AC, green = CAR, blue = fiber head).

Mentions: We found that the sialic acid binding site on CAV-2 head is distinct, both in sequence and location, from that found on the HAd37 fiber head. On the CAV-2 fibers head, sialic acid binds further away from the three-fold symmetry axis at the periphery of the trimer (Figure 3A–D). The residues involved in binding (Asn435, Ser419, Ser416, Gln417 and Arg515) do not align with those involved in sialic acid-binding by HAd37 fiber head (Figure S2). The HAd37 sialic acid binding site consists of three residues forming hydrogen bonds (Tyr213, Pro317 and Lys345) and two residues contacting sialic acid in hydrophobic interactions (Tyr308 and Val322) (Figure 3E). All seven interactions between CAV-2 fiber head and sialic acid are hydrogen bonds or salt bridges (Figure 3F), suggesting a relatively strong interaction compared to HAd37. Unlike HAd37 fiber head [33], no hydrophobic contacts contribute to sialic acid binding. Finally, sialic acid binds within a basic patch in each head (Figure 3C and D).


The cell adhesion molecule "CAR" and sialic acid on human erythrocytes influence adenovirus in vivo biodistribution.

Seiradake E, Henaff D, Wodrich H, Billet O, Perreau M, Hippert C, Mennechet F, Schoehn G, Lortat-Jacob H, Dreja H, Ibanes S, Kalatzis V, Wang JP, Finberg RW, Cusack S, Kremer EJ - PLoS Pathog. (2009)

Structure of the CAV-2 fiber head in complex with sialic acid and CAR.A. Ribbon diagram of CAV-2 fiber head in complex with sialyl-D-lactose (red = Neu5AC, yellow = fiber head). B. Cartoon representation of HAd37 fiber head in complex with sialyl-D-lactose (red = sialic acid, blue = fiber head). C. Surface charge representation of CAV-2 fiber head and cartoon representation of sialic acid as found in the complex. D. Surface charge representation of HAd37 fiber head and cartoon representation of sialic acid as found in the complex. E. Representation of the contacts between CAV-2 fiber head (purple) and sialic acid (yellow). F. Representation of the contacts between HAd37 fiber head (purple) and Neu5AC (yellow). G. Ribbon diagram of CAV-2 fiber head in complex with CAR D1 and sialyl-D-lactose (red = Neu5AC, green = CAR, yellow = fiber head). H. Ribbon diagram of HAd37 fiber head in complex with CAR and sialyl-D-lactose (red = Neu5AC, green = CAR, blue = fiber head).
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ppat-1000277-g003: Structure of the CAV-2 fiber head in complex with sialic acid and CAR.A. Ribbon diagram of CAV-2 fiber head in complex with sialyl-D-lactose (red = Neu5AC, yellow = fiber head). B. Cartoon representation of HAd37 fiber head in complex with sialyl-D-lactose (red = sialic acid, blue = fiber head). C. Surface charge representation of CAV-2 fiber head and cartoon representation of sialic acid as found in the complex. D. Surface charge representation of HAd37 fiber head and cartoon representation of sialic acid as found in the complex. E. Representation of the contacts between CAV-2 fiber head (purple) and sialic acid (yellow). F. Representation of the contacts between HAd37 fiber head (purple) and Neu5AC (yellow). G. Ribbon diagram of CAV-2 fiber head in complex with CAR D1 and sialyl-D-lactose (red = Neu5AC, green = CAR, yellow = fiber head). H. Ribbon diagram of HAd37 fiber head in complex with CAR and sialyl-D-lactose (red = Neu5AC, green = CAR, blue = fiber head).
Mentions: We found that the sialic acid binding site on CAV-2 head is distinct, both in sequence and location, from that found on the HAd37 fiber head. On the CAV-2 fibers head, sialic acid binds further away from the three-fold symmetry axis at the periphery of the trimer (Figure 3A–D). The residues involved in binding (Asn435, Ser419, Ser416, Gln417 and Arg515) do not align with those involved in sialic acid-binding by HAd37 fiber head (Figure S2). The HAd37 sialic acid binding site consists of three residues forming hydrogen bonds (Tyr213, Pro317 and Lys345) and two residues contacting sialic acid in hydrophobic interactions (Tyr308 and Val322) (Figure 3E). All seven interactions between CAV-2 fiber head and sialic acid are hydrogen bonds or salt bridges (Figure 3F), suggesting a relatively strong interaction compared to HAd37. Unlike HAd37 fiber head [33], no hydrophobic contacts contribute to sialic acid binding. Finally, sialic acid binds within a basic patch in each head (Figure 3C and D).

Bottom Line: Using molecular, biochemical, structural and transgenic animal-based analyses, we found that the primary erythrocyte interaction domain for HAd37 is its sialic acid binding site, while CAV-2 binding depends on at least three factors: electrostatic interactions, sialic acid binding and, unexpectedly, binding to the coxsackievirus and adenovirus receptor (CAR) on human erythrocytes.We show that the presence of CAR on erythrocytes leads to prolonged in vivo blood half-life and significantly reduced liver infection when a CAR-tropic Ad is injected intravenously.This study provides i) a molecular and structural rationale for Ad-erythrocyte interactions, ii) a basis to improve vector-mediated gene transfer and iii) a mechanism that may explain the biodistribution and pathogenic inconsistencies found between human and animal models.

View Article: PubMed Central - PubMed

Affiliation: European Molecular Biology Laboratory, Grenoble Outstation, Grenoble, France.

ABSTRACT
Although it has been known for 50 years that adenoviruses (Ads) interact with erythrocytes ex vivo, the molecular and structural basis for this interaction, which has been serendipitously exploited for diagnostic tests, is unknown. In this study, we characterized the interaction between erythrocytes and unrelated Ad serotypes, human 5 (HAd5) and 37 (HAd37), and canine 2 (CAV-2). While these serotypes agglutinate human erythrocytes, they use different receptors, have different tropisms and/or infect different species. Using molecular, biochemical, structural and transgenic animal-based analyses, we found that the primary erythrocyte interaction domain for HAd37 is its sialic acid binding site, while CAV-2 binding depends on at least three factors: electrostatic interactions, sialic acid binding and, unexpectedly, binding to the coxsackievirus and adenovirus receptor (CAR) on human erythrocytes. We show that the presence of CAR on erythrocytes leads to prolonged in vivo blood half-life and significantly reduced liver infection when a CAR-tropic Ad is injected intravenously. This study provides i) a molecular and structural rationale for Ad-erythrocyte interactions, ii) a basis to improve vector-mediated gene transfer and iii) a mechanism that may explain the biodistribution and pathogenic inconsistencies found between human and animal models.

Show MeSH
Related in: MedlinePlus